Southern blotting

A method developed by EM Southern for detecting and manipulating specific DNA sequences previously separated by gel electrophoresis using a radioactively- or chemically-labeled DNA probe with sequence similarity

Southern blotting

A method of identifying a fragment of DNA containing a specific sequence of bases. A mixture of fragments, produced by cutting DNA with restriction enzymes, is separated on a gel block by ELECTROPHORESIS. The fragments are denatured to single strand DNA and transferred, by blotting, to a nitrocellulose sheet. The position of the desired fragment can then be shown by hybridizing it with a DNA probe labelled with radioactive phosphorous that will cause a black line on an X-ray film. The method was named after its developer Edward M. Southern. A similar technique for RNA analysis has been humorously called ‘Northern blotting’ and the play on words has been extended to include ‘WESTERN BLOTTING’.

Predicted restriction enzyme cut sites in the database sequences were compared with actual restriction mapping fragment sizes and with the data from Southern blotting to determine the order of sequence contigs, An independent sequence assembly for this region obtained from Celera Genomics (GA_x6K02T2PU9B, as of 1 June 2002) was also used to fill in some gaps.

The Sequenom EpiTYPER system (which uses MALDI-TOF mass spectrometry for quantification) and Southern blotting tools were used to determine the methylation output ratio and the activation ratio, respectively.

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